Purification And Characterization Of Superoxide Dismutase From Martianus Dermestoides Chevrola

Superoxide dismutase(SOD)is a metal-containing enzyme which catalyzes the dismutation reaction.It has been found that it is relational with the organism anti-decrepit,anti-tumor, nutrition situations,immunity disease,the inflammation and protection.The adult of Martianus dermestoides Chevrolat has high economic value for its abundant functional ingredients,such as necessary amino acids,vitamin,microelements and protein,unsaturated fatty acids et al.This paper attempted to make a research on Martianus dermestoides Chevrolat. and it was focused on extraction,purification and characterization of superoxide dismutase from Martianus dermestoides Chevrolat.In this paper,the extraction conditions of superoxide dismutase(SOD)from Martianus dermestoides was optimized by one factor as well as orthogonal experimental design.The result showed that SOD extracted using NaCl had a higher activity compared with the conventional phosphate extraction method.The concentration of extraction solution had the greatest effect on SOD extraction.The optimal extraction conditions for SOD from Martianus dermestoides are using 0.13 M NaCl with a 1:13 ratio to extract at 28℃for 14 h.The activity of SOD was 38.83U/ml,and the specific enzymatic activity of SOD was 123.66U/mg pro. During the extraction process,most of the unwanted protein can be removed though a heat-denatured condition at 60,for 15min.The SOD was purified by the following methods:dialysis,polyethylene glycol 20000 (PEG20000)concentration,Sephadex G-75 gel chromatography(1.6×50 cm),DEAE-Sepharose FF(1.6×50cm)ion exchange chromatography.The result showed that the purification fold was 3.86 and the activity yield was 21.89%.It is a sole protein belt and the molecular weight is 40.58Kda.The effects of H₂O₂ can obviously inhibit the enzyme,and CHCl₃-CH₃CH₂OH only demonstrated a slight inhibitory effect,this enzyme was proved as Cu/Zn-SODFollowing purification,some enzymatic characterizations of the SOD were studied.The optimum reaction temperature of purified SOD was 50℃.The optimum reaction pH value of purified was 6.It is stable below 50℃and at pH 5-8...